FIG. 3.

Effect of confluence on surface expression of NG2 and CD146, proliferation potential and potency. Pooled P2 OAC cultures from five single cell-derived colonies were inoculated at 100 cells/cm² into T-150 flasks containing 20 mL medium. Preconfluent cultures were analyzed 5 days after inoculation. T-flask cultures that were confluent for 7 days were passaged once to generate postconfluent samples, which were analyzed 5 days after passaging when the cells were <50% confluent. Surface expression of NG2 (A) and CD146 (B) reported as an MFI ratio as in Figure 1. Proliferation potential was evaluated by monitoring (C) cell doublings during the first 5 days after inoculation and (D) colony-forming efficiency when cell samples were plated at clonogenic levels. Cell potency to exhibit osteo- (E), adipo-, (F) and chondrogenesis (G) was quantified as in Supplementary Figure S1. Data are expressed as a mean±standard error of the mean (n=3). *p<0.05 and **p<0.01 versus preconfluent culture.